The liver neutralises microbial infections and detoxifies xenobiotics. However, exposure to these agents results in liver cell damage which necessitates a rapid and efficient wound-healing response. Central to this wound-healing is the local production of scar-forming myofibroblasts. A rapid response mechanism for generating hepatic myofibroblasts is the transdifferentiation of resident quiescent retinoid-storing hepatic stellate cells (Friedman S L 2008). Myofibroblast transdifferentiation (MTD) also occurs with pancreatic stellate cells and renal mesangial cells in the injured pancreas and kidney respectively (Ornery et al JCI-2007, Simonson M S 2007), suggesting biological conservation of the process. MTD is associated with global changes in gene transcription required for the cell to adopt the pro-inflammatory and pro-fibrogenic characteristics of the myofibroblast (Smart and Mann 2002). Regulation of MTD-associated gene expression is poorly understood but must be under strict control to prevent inappropriate wound healing (or fibrosis). Here we describe a novel epigenetic relay that is initiated by loss of expression of microRNA miR132 and which culminates in transcriptional silencing of PPARγ, a master negative regulator of MTD of hepatic stellate cells (She H et al 2005, Tsukamoto H et al 2006). We further describe two key components of the relay pathway, MeCP2 and EZH2, as critical regulators of hepatic wound-healing.
The present invention relates to the use of a compound according to the chemical structure Ia:
Where B is
W is C—H, O or S (preferably C—H or O, more preferably C—H) such that the bond between W and the adjacent carbon atom is a double bond when W is C—H and a single bond when W is O or S;V is C-A″, O or S, preferably with the proviso that when V is O or S, W is O or S (preferably, both V and W are O);A is H, OR2 or halogen (F, Cl, Br, I, preferably F or Br, more preferably F);A′ is H, OR2 or halogen (F, Cl, Br, I, preferably F or Br, more preferably F);A″ is H or OR1, with the proviso that when A′ is OR2, A is H; and when A is OR2, A′ is H;X is C—R3 or N;Y is C—R3 or N; preferably X or Y is N and X and Y are not both simultaneously N;Rz is H or a C1-C3 alkyl group, optionally substituted with OH (preferably H);R3 is H, a halogen or C1-C3 alkyl;D is H, a halogen (preferably F, Cl or Br) or NR1aR2;E is absent (when G is NHR2) or H (when G is O);G is O or NR1aR2;J is N or C—R4;K is N or C—H;R4 is H, halogen (F, Cl, Br, I), CN, —C(═O)NH2, NH2, NO2, —C═C—H (cis or trans) or —C≡C—H;Ra is H or CH3;Each R1 is independently H, an acyl group, a C1-C20 alkyl or ether group, an amino acid (D or L), a phosphate, diphosphate, triphosphate, phosphodiester group;Each R1a and R2 is independently H, an acyl group, a C1-C20 alkyl or ether group, an amino acid (D or L) or together R1a and R2 form a C3-C7 cycloalkyl group; andpharmaceutically acceptable salts, solvates or polymorphs thereof to treat and/or inhibit fibrogenesis in a patient or subject especially including fibrotic disease and/or conditions, including liver fibrosis and/or cirrhosis of the liver, in particular fibrosis and cirrhosis which may be caused by viruses, chemical and/or drugs.
Certain alternative embodiments for use in treating and/or inhibiting fibrogenesis, including fibrotic disease and/or conditions as otherwise described herein include compounds wherein W and V are both O and wherein B, A, A′ and R1 are the same as described for formula Ia above.
Alternative preferred compounds for use in treating and/or inhibiting fibrogenesis, including fibrotic disease and/or conditions as otherwise described herein include compounds according to the chemical structure Ib (W is a C—H):
Wherein B, A, A′, A″ and R1 are the same as described for formula Ia above.
Fibrotic diseases which may be treated according to the present invention include, for example, liver fibrosis (alcoholic, viral, autoimmune, metabolic and hereditary chronic disease), renal fibrosis (e.g., resulting from chronic inflammation, infections or type II diabetes), lung fibrosis (idiopathic or resulting from environmental insults including toxic particles, sarcoidosis, asbestosis, hypersensitivity pneumonitis, bacterial infections including tuberculosis, medicines, etc.), interstitial fibrosis, systemic scleroderma (autoimmune disease in which many organs become fibrotic), macular degeneration (fibrotic disease of the eye), pancreatic fibrosis (resulting from, for example, alcohol abuse and chronic inflammatory disease of the pancreas), fibrosis of the spleen (from sickle cell anemia, other blood disorders) cardiac fibrosis (resulting from infection, inflammation and hypertrophy), mediastinal fibrosis, myelofibrosis, endomyocardial fibrosis, retroperitoneal fibrosis, progressive massive fibrosis, nephrogenic systemic fibrosis, fibrotic complications of surgery, especially surgical implants, injection fibrosis and secondary conditions and disease states of fibrosis. Secondary conditions and disease states of fibrosis include for example, cirrhosis, diffuse parenchymal lung disease, post-vasectomy pain syndrome and rheumatoid arthritis, among others.
In certain preferred aspects of the present invention (especially including compounds according to formula Ib), A is OH, A′ is H and A″ is OH, J is N or CR4, K is N or CH, X is N, Y is CR3, E is absent and G is NHR2. In other preferred embodiments, J is N, K is CH and G is O or NHR2. In many preferred embodiments, R1 and R2 are both H. In certain preferred embodiments, R4 is an acetylenic group.
In other embodiments, the preferred compound is
Where R1, R2, R4, X and Y are the same as described above. Other preferred compounds may be readily gleaned from the description of the invention which follows.
In still other preferred embodiments, the compound is according to the chemical structure Ic hereinbelow:
Where B′ is
Or pharmaceutically acceptable salts, solvates or polymorphs thereof.
In still other preferred embodiments compounds which may be used in the present invention include the following:
Where D is H, F, Cl or Br, preferably F or Cl, more preferably Cl,Or pharmaceutically acceptable salts, solvates or polymorphs thereof.
In alternative embodiments, the compound as described hereinabove, may be conjugated to an antibody (monoclonal or polyclonal) which binds to MeCP2 or EZH2 (anti-MECP2 or anti-EZH2). Alternatively, the above antibodies may be used in the absence of conjugation to inhibit MeCP2 or EZH2 in order to inhibit fibrogenesis in a patient or subject, including liver fibrogenesis and to treat cirrhosis of the liver, as well as fibrotic disease and disease states and conditions which occur secondary to fibrogenesis and/or fibrotic disease.
The present invention also relates to pharmaceutical compositions comprising an effective amount of any one or more of the compounds described above (especially compounds conjugated to mono and/or polyclonal antibodies), optionally in combination with a pharmaceutically acceptable carrier, additive or excipient.
Thus, the present application is directed to the inhibition of fibrogenesis, including liver fibrogenesis in a patient in need thereof, and/or the treatment of cirrhosis of the liver (which may be caused for example, by a virus, a chemical or drug) comprising administering an effective amount of one or more compounds or anti-MeCP2 and/or anti-EZH2 antibodies (including compounds conjugated to anti-MECP2 and/or anti-EZH2 antibodies) according to the present invention optionally in combination with a pharmaceutically acceptable carrier, additive or excipient to said patient. Pharmaceutical compositions based upon these antibodies and/or nucleoside compounds in effective amounts in combination with a pharmaceutically acceptable carrier, additive or excipient are additional aspects of the present invention.
The present invention also relates to the inhibition of methylation of DNA and RNA in cells comprising exposing cells, especially including liver cells to an effective of a compound as otherwise disclosed herein. A method of inhibiting the methylation of DNA and/or RNA in cells, especially liver cells, in a patient comprises administering an effective amount of a compound as otherwise described herein to said patient.
The present invention also relates to method for inhibiting and/or treating fibrotic diseases including, for example, liver fibrosis (alcoholic, viral, autoimmune, metabolic and hereditary chronic disease), renal fibrosis (e.g., resulting from chronic inflammation, infections or type II diabetes), lung fibrosis (idiopathic or resulting from environmental insults including toxic particles, sarcoidosis, asbestosis, hypersensitivity pneumonitis, bacterial infections including tuberculosis, medicines, etc.), interstitial fibrosis, systemic scleroderma (autoimmune disease in which many organs become fibrotic), macular degeneration (fibrotic disease of the eye), pancreatic fibrosis (resulting from, for example, alcohol abuse and chronic inflammatory disease of the pancreas), fibrosis of the spleen (from sickle cell anemia, other blood disorders) cardiac fibrosis (resulting from infection, inflammation and hypertrophy), mediastinal fibrosis, myelofibrosis, endomyocardial fibrosis, retroperitoneal fibrosis, progressive massive fibrosis, nephrogenic systemic fibrosis, fibrotic complications of surgery, especially surgical implants, injection fibrosis and secondary conditions and disease states of fibrosis. Secondary conditions and disease states which occur as a consequence of or associated with fibrosis include for example, cirrhosis, diffuse parenchymal lung disease, post-vasectomy pain syndrome and rheumatoid arthritis, among others. The method according to the present invention comprises administering an effective amount of one or more compounds according to the present invention to a patient at risk for a fibrotic disease or in need of therapy for a fibrotic disease or secondary disease state or condition thereof, optionally in combination with a pharmaceutically acceptable carrier, additive or excipient.